CN1071458A - Be rich in the alcohol fermentation method of the spentsulfiteliquor of pentose - Google Patents
Be rich in the alcohol fermentation method of the spentsulfiteliquor of pentose Download PDFInfo
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- CN1071458A CN1071458A CN 92107774 CN92107774A CN1071458A CN 1071458 A CN1071458 A CN 1071458A CN 92107774 CN92107774 CN 92107774 CN 92107774 A CN92107774 A CN 92107774A CN 1071458 A CN1071458 A CN 1071458A
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- spentsulfiteliquor
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- nutrient solution
- pentose
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Abstract
A kind of method of producing alcohol after sulphite paper-making pulping waste liquid is fermented with microorganism is characterized in that with shehatae candida (Candida shehatae) be organism of fermentation, and during the fermentation to fermentor tank little air without interruption.Use method provided by the present invention, the spentsulfiteliquor that is rich in pentose is after taming and dociling the shehatae candida fermentation of educating, and the utilization ratio of sugar (comprising pentose and hexose) can reach 90%, and spirit yield can reach 0.40 gram
Alcohol/ gram
SugarMore than.
Description
The invention belongs to microbial fermentation and produce the alcohol technology.
Spentsulfiteliquor is that timber or other lignocellulose raw material (as reed, agricultural crop straw etc.) are isolated remaining brown liquid behind the man-made fiber paper pulp through the sulphite process high temperature steaming in the paper industry.Mainly contain sulfonated lignin, sugar (comprising hexose and pentose), acid, aldehyde material, sulfurous gas and inorganic salt etc.
Utilize sugar-fermenting in the spentsulfiteliquor to produce the technology of alcohol, the industrial application history of decades has been arranged.Fermenting speed is fast, adaptive faculty by force, does not need advantages such as oxygen supply owing to having for yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), is commonly industry member and adopts.But yeast saccharomyces cerevisiae can only utilize the hexose (comprising glucose, seminose and semi-lactosi) in the waste liquid, the pentose (comprising wood sugar and pectinose) in can't the metabolism waste liquid.
Piperylene is one of main ingredient of timber and other lignocellulose raw material hemicellulose.In softwood, piperylene accounts for 5～10% of timber dry weight; In pourous wood and agricultural crop straw, pentosan content can be up to 15～30% of raw material dry weight.In the sulphite pulping process, piperylene is hydrolyzed into and is pentose.In recent years, along with the minimizing of China's forest zone softwood resource, pourous wood and agricultural crop straw are more and more higher with the ratio in the material in sulphite slurrying.This spentsulfiteliquor that is rich in pentose ferments with traditional yeast saccharomyces cerevisiae, because the very high pentose of content in can't the metabolism waste liquid, so spirit yield is very low, the economic benefit extreme difference.
Whether effective utilization of pentose also has influence on the comprehensive utilization of another main ingredient-sulfonated lignin in the spentsulfiteliquor.Sulfonated lignin are widely used as tackiness agent, dispersion agent, cement water reducing agent, oil drilling slurry thinner etc.In most of the cases, the user requires the sugar degree in the lignosulfonic acid product salt to be lower than 5%.If can utilize the pentose fermentation technology that pentose in the waste liquid and hexose are changed into alcohol synchronously, then not only can increase alcohol output, and can guarantee that the sugar degree of lignosulfonic acid product salt is lower than 5%, helps the comprehensive utilization of waste liquid greatly.
The zymamsis technology of pentose is the emerging research field of the eighties in the world.For a long time, scientific circles think always that pentose can not be fermented effectively by microorganism becomes alcohol.The oil crisis of late nineteen seventies, early eighties has caused from reproducible agriculture and forestry plant fibre resources produces the Study of Alcohol upsurge, and pentose fermentation has been subjected to the great attention of countries in the world as one of gordian technique of this research.Through effort about ten years, the research in this field has obtained many impressive progresses in the world, relates to the document emerge in multitude of pentose fermentation bacterial classification, fermenting mechanism.But up to now, Shang Weiyou is applicable to the technology report of the spentsulfiteliquor zymamsis of being rich in pentose, and its major cause is:
1. the chemical composition complexity of spentsulfiteliquor, the acetic acid of the content height, particularly high density of fermentation inhibitor and sulfurous gas have serious inhibition to the metabolism of pentose;
2. pentose fermentation need be supplied with minor amounts of oxygen, and the fermentation of hexose then needs under anaerobic to carry out, and the balance of the two has many difficulties;
3. in the fermenting process, the hexose in the waste liquid is preferentially utilized by microorganism, and its metabolic intermediate has restraining effect to the utilization of pentose.
The objective of the invention is to seek the alcohol fermentation method that is applicable to the spentsulfiteliquor that is rich in pentose, comprise bacterial classification selection, tame and docile and to educate the pre-treatment of waste liquid, technological condition for fermentation etc.
The microorganism that the present invention is used to ferment is chosen as shehatae candida (Candida shehatae).This zymic starting strain ATCC 22984 is by USS strain library (American Type Culture Collection, Rockville, Maryland, U.S.A.) public sale.ATCC 22984 has the ability that pentose is converted into alcohol, but poor to the adaptive faculty of spentsulfiteliquor, need tame and docile and educate.
Bacterial classification tame and docile method of educating in spentsulfiteliquor is identical with the tame and docile method of educating of traditional yeast saccharomyces cerevisiae basically.Promptly under identical condition, make yeast growth many generations, progressively change the ratio that substratum is formed then.The substratum that is adopted is formed the change rule: from nutritious wood sugar nutrient solution, reduce the amount of wood sugar nutrient solution gradually, and corresponding increase spentsulfiteliquor nutrient solution, until yeast is cultivated in 100% spentsulfiteliquor nutrient solution.Every change one subculture is formed, and carries out the fermentation comparison test of certain number of times, and after obtaining satisfied result, the bacterial classification that enters again after next round change substratum is formed is many for grown cultures.
Wood sugar nutrient solution of the present invention is made up of wood sugar 20～30g/L, peptone 3～5g/L, Yeast diffusion juice 1～3g/L, natural pH.
Educate through long-term the taming and dociling in spentsulfiteliquor, important variation has taken place in the form of shehatae candida and physio-biochemical characteristics.Compare with starting strain ATCC 22984, become big through taming and dociling strain cell volume after educating, cell walls thickens, content of soluble protein increases in the cell, quantity increases, vigor strengthens to help isozyme (as alcohol dehydrogenase, zymohexase, hexokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, malate dehydrogenase (malic acid dehydrogenase), isocitric enzyme etc.) that pentose is converted into alcohol, all increases substantially to the adaptive faculty of spentsulfiteliquor and to the fermentation capacity of pentose.And through tame and docile bacterial strain stable performance of educating, irreversible.
Adopt to tame and docile shehatae candida after educating and promptly can be used for being rich in the zymamsis of the spentsulfiteliquor of pentose.But must carry out pre-treatment to spentsulfiteliquor before the fermentation.Pretreated method is: with spentsulfiteliquor under 80～85 ℃ of temperature, slowly add lime powder or milk of lime while stirring, in and waste liquid in contained acid, pH is 5.3～5.8(pourous wood pulping waste liquor ratio capping when higher after being cooled to room temperature, takes off limit when softwood pulping waste liquor ratio is higher).Leave standstill clarification, discard sediment, get its clear liquid and use for fermentation.
Get above-mentioned shehatae candida and spentsulfiteliquor pretreatment fluid of taming and dociling after educating, the ratio that adds dry yeast cell 3～5 grams with every liter of pretreatment fluid, add nitrogen salts such as quantitative urea or ammonium sulfate, ammonium nitrate, Secondary ammonium phosphate, ammoniacal liquor again, and microcosmic salts such as phosphoric acid or calcium superphosphate are nutritive salt, putting into the batch fermentation jar or the jar that continuously ferments, is 30～38 ℃, dissolved oxygen coefficient K in temperature
LA is the processing condition bottom fermentation 15～36 hours of 80～150l/h.Can distill according to a conventional method after fermentation is finished and obtain the finished product industrial spirit.
The description of the drawings:
Fig. 1 adopts method of the present invention, in the alcoholic fermentation process of the spentsulfiteliquor that is rich in pentose, and sugar, alcohol, yeast cell and the time dependent typical law of pH value.Among Fig. 1:
Curve 1 expression ethanol concn;
Curve 2 expression barm cell concentrations;
Curve 3 expression pH values;
Curve 4 expression total sugar contents.
Embodiment one, and the tame and docile of shehatae candida educates:
1. the wood sugar nutrient solution is made up of wood sugar 20～30g/L, peptone 3～5g/L, Yeast diffusion juice 1～3g/L, natural pH;
2. spentsulfiteliquor slowly adds lime powder or milk of lime while stirring under 80～85 ℃ of temperature, in and waste liquid in contained acid, pH is 5.3～5.8 after being cooled to room temperature.Leave standstill clarification, discard sediment, get its clear liquid, add peptone 0.3～0.5g/L, Yeast diffusion juice 0.1～0.3g/L, make the pulping waste liquor nutrient solution.
Step 1:
Containing 95ml wood sugar nutrient solution and 5ml pretreatment fluid in volume is in the triangular flask of 250ml, band cotton plug, and placing temperature is that 38 ℃, revolution shake speed and are the air constant temperature oscillator shaking culture of 150r/min 24～48 hours.Whenever be commissioned to train support to finish after, with nutrient solution centrifugal treating under the centrifugal acceleration of 500～1500g, get the yeast cell precipitation and insert in the mixed-culture medium of the same ratio of future generation, repeat said process, cultivate (result with the fermentation comparison test is a foundation) about 10 generations.
Step 2:
Gained yeast cell precipitation after the centrifugal treating in the step 1 is inserted in the mixed-culture medium of being made up of 90ml wood sugar nutrient solution and 10ml pretreatment fluid the culturing process of repeating step one.
Later on the component of each mixed-culture medium changes to be and reduces 5ml wood sugar nutrient solution and increase the 5ml pretreatment fluid, until nutrient solution consist of 100% spentsulfiteliquor pretreatment fluid till.
Embodiment two, the pretreatment fluid preparation of spentsulfiteliquor:
Softwood (Korean pine, kahikatea etc.) pulping waste liquor 30%; Pourous wood (poplar etc.) pulping waste liquor 70%, when mixed pulp waste liquid atmospheric evaporation is concentrated into solid content and is 15% left and right sides, under 80～85 ℃ of temperature, slowly add lime powder or milk of lime while stirring, in and waste liquid in contained acid, pH is 5.3～5.5 after being cooled to room temperature.Leave standstill clarification, discard sediment, get its clear liquid and use for fermentation.
Embodiment three, the preparation of the pretreatment fluid of spentsulfiteliquor:
When softwood (Pinus massoniana Lamb) pulping waste liquor atmospheric evaporation is concentrated into solid content and is 13% left and right sides, under 80～85 ℃ of temperature, slowly add lime powder or milk of lime while stirring, in and waste liquid in contained acid, pH is 5.3 after being cooled to room temperature.Leave standstill clarification, discard sediment, get its clear liquid and use for fermentation.
Embodiment four, the preparation of the pretreatment fluid of spentsulfiteliquor:
When reed pulping waste liquor atmospheric evaporation is concentrated into solid content and is 13% left and right sides, under 80～85 ℃ of temperature, slowly add lime powder or milk of lime while stirring, in and contained acid in the waste liquid, pH is 5.8 after being cooled to room temperature.Leave standstill clarification, discard sediment, get its clear liquid and use for fermentation.
Embodiment five, the zymotechnique of batch fermentation jar:
1. fermenation raw liquid adopts among the embodiment two by softwood (Korean pine, kahikatea etc.) pulping waste liquor 30%; The pretreatment fluid of pourous wood (poplar etc.) pulping waste liquor 70% preparation.
2. the add-on of nutritive salt is: urea 0.24 grams per liter;
Phosphoric acid 0.05 grams per liter.
3. taming and dociling the shehatae candida add-on of educating by embodiment one is 4.0 grams per liters.
The fermented liquid temperature control is made as 38 ± 1 ℃, is 2.51/l with flow
Fermented liquidThe air of h is imported the core gas distributor bubbling in the fermentor tank continuously equably, and the agitator with per minute 150r/min stirs fermented liquid again.Fermented 34 hours.
That measures in the fermenting process the results are shown in Table 1.
The zymotechnique that six, three grades of series connection of embodiment are continuously fermented jar:
Three grades of series connection continuously ferment device by the stoste storage tank, (every jar of liquid capacity is 0.4m to three fermentor tanks
3), yeast separation machine, volumetric infusion pump and air purifier form.Each fermentor tank disposes straight turbine agitator, air distribution pipe and the hot water or cold water's heat exchange coiled pipe of taking turns of six leaves that rotating speed is 250r/min.Fermenation raw liquid is with continuous No. 1 fermentor tank of input of volumetric infusion pump, by the upflow tube fermentor tank of progressively flowing through No. 2, No. 3, enters dish formula nozzle deslagging yeast whizzer at last and separates.The yeast concentrated emulsion that separation obtains is back to No. 1 fermentor tank and recycles.Pressurized air feeds each fermentor tank and carries out bubbling after the efficient metal strainer filters.
The processing condition that adopt are as follows:
1. fermenation raw liquid adopts the pretreatment fluid for preparing with softwood (Pinus massoniana Lamb) pulping waste liquor among the embodiment three.
2. the add-on of nutritive salt is: urea 0.24 grams per liter;
Phosphoric acid 0.05 grams per liter.
3. taming and dociling the shehatae candida add-on of educating by embodiment one is 4.0 grams per liters.
The fermented liquid temperature control is made as 38 ± 1 ℃, and air feeding amount is:
No. 1 fermentor tank 0.5m
3/ h
No. 2 fermentor tank 0.4m
3/ h
No. 3 fermentor tank 0.4m
3/ h
The input of fermenation raw liquid is 0.05m
3/ h; The quantity of reflux of yeast concentrated emulsion is 0.01m
3/ h.
That measures in the fermenting process the results are shown in Table 2.
Embodiment seven:
Appointed condition is with embodiment five.The processing condition that adopt are as follows:
1. fermenation raw liquid adopts the pretreatment fluid of reed pulping waste liquor preparation among the embodiment four.
2. the add-on of nutritive salt is: urea 0.16 grams per liter;
Phosphoric acid 0.03 grams per liter.
3. taming and dociling the shehatae candida add-on of educating by embodiment one is 5.0 grams per liters.
The fermented liquid temperature control is made as 32 ± 1 ℃, is 1.51/l with flow
Fermented liquidThe air of h is imported the core gas distributor bubbling in the fermentor tank continuously equably, and the agitator with per minute 500r/min stirs fermented liquid again.Fermented 24 hours.
That measures in the fermenting process the results are shown in Table 3.
Use method provided by the present invention, the spentsulfiteliquor that is rich in pentose is after taming and dociling the shehatae candida fermentation of educating, and the utilization ratio of sugar (comprising pentose and hexose) can reach 90%, and spirit yield can reach 0.40 gram
Alcohol/ gram
SugarMore than, be more than 80% of theoretical yield.From accompanying drawing 1, table 1, table 2 and table 3 as can be seen, the change in concentration rule of sugar meets the first order reaction rule in the fermenting process, explanation is under the pentose content condition with higher, although hexose still preferentially obtains utilizing, the utilization to pentose does not produce the obvious suppression effect.The generation of alcohol is carried out basically synchronously with the utilization of sugar, illustrates that the accumulation of intermediate product is not obvious.Because the air feed rate is more reasonable, barm cell concentration remains unchanged in whole fermentation process substantially.Present method is compared with the fermentation by saccharomyces cerevisiae method of industrial widespread use, and the utilization ratio and the alcohol quantum of output of sugar have all improved 20～140%.In the sulfonated lignin dry powder that waste liquid spraying drying after the fermentation obtains, the content of sugar is reduced to below 3%, and is more much lower than the sugar degree (10～15%) of conventional gained dry powder, helps the further utilization of sulfonated lignin dry powder.Therefore, present method has tangible society, economic benefit.
Claims (4)
1, a kind of method of producing alcohol after sulphite paper-making pulping waste liquid is fermented with microorganism is characterized in that:
A. organism of fermentation is shehatae candida (Candida shehatae);
B. must be in the fermenting process to fermentor tank little air without interruption.
2, the method for producing alcohol as claimed in claim 1 is characterized in that the tame and docile process of educating of shehatae candida is:
A. spentsulfiteliquor is added lime powder or milk of lime, being neutralized to pH is 5.3～5.8.Leave standstill clarification, discard sediment, add peptone, Yeast diffusion juice after getting its clear liquid, make the pulping waste liquor nutrient solution, a small amount of pulping waste liquor nutrient solution is added in the wood sugar nutrient solution of being made up of wood sugar, peptone, Yeast diffusion juice make mixed-culture medium;
B. mixed-culture medium is contained in container, placed air constant temperature oscillator shaking culture.Whenever be commissioned to train and medium centrifugal handled after support finishing, get the yeast cell precipitation and insert in the mixed-culture medium of the same ratio of future generation, repeat said process, be cultured to leavening property reliable and stable till.
C. increase the ratio of pulping waste liquor nutrient solution in the mixed-culture medium on a small quantity, repetitive process b, until nutrient solution consist of 100% spentsulfiteliquor nutrient solution till.
3, the method for producing alcohol as claimed in claim 1 is characterized in that spentsulfiteliquor needs through following pre-treatment before fermentation:
When the pulping waste liquor atmospheric evaporation is concentrated into solid content and is 13～15% left and right sides, under 80～85 ℃ of temperature, slowly add lime powder or milk of lime while stirring, in and contained acid in the waste liquid, pH is 5.3～5.8 after being cooled to room temperature.Leave standstill clarification, discard sediment, get its clear liquid and use for fermentation.
4, the method for producing alcohol as claimed in claim 1 is characterized in that spentsulfiteliquor need add a small amount of nitrogen salt and microcosmic salt when fermentation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92107774 CN1035390C (en) | 1992-09-28 | 1992-09-28 | Fermentation method for producing alcohol with pentose-containing sulfite pulping waste liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92107774 CN1035390C (en) | 1992-09-28 | 1992-09-28 | Fermentation method for producing alcohol with pentose-containing sulfite pulping waste liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1071458A true CN1071458A (en) | 1993-04-28 |
| CN1035390C CN1035390C (en) | 1997-07-09 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 92107774 Expired - Fee Related CN1035390C (en) | 1992-09-28 | 1992-09-28 | Fermentation method for producing alcohol with pentose-containing sulfite pulping waste liquid |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112125563A (en) * | 2020-09-24 | 2020-12-25 | 张百顺 | Production method of concrete water reducing agent |
-
1992
- 1992-09-28 CN CN 92107774 patent/CN1035390C/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112125563A (en) * | 2020-09-24 | 2020-12-25 | 张百顺 | Production method of concrete water reducing agent |
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| CN1035390C (en) | 1997-07-09 |
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